Flavopereirine and alstonine combinations in the treatment and prevention of prostate cancer

ABSTRACT

A method and composition for preventing prostate cancer and/or reducing PSA levels and/or alleviating the symptoms of BPH (Benign Prostatic Hyperplasia) or PIN (prostatic intraepithelial neoplasia) by administration of an effective amount of a mixture of flavopereirine and alstonine. A method and composition for treating low-grade prostate cancer and preventing the onset of metastatic disease and/or reducing the doubling time of PSA levels in men with positive biopsies showing relatively low Gleason scores and morphologies characteristic of non-invasive, slow-progressing prostate cancer. The flavopereirine and alstonine can be in the form of natural extracts derived from  Pao Pereira  and  Rauwolfia Vomitoria , respectively. Alternatively, these two active compounds can be administered in purified form. The composition can be in included in a kit along with instructions for use in a treatment regimen.

FIELD OF INVENTION

This invention is in the field of the prevention and treatment ofprostate cancer. More specifically, this invention relates to the use offlavopereirine and alstonine combinations in preventing and treatingprostate cancer.

BACKGROUND OF THE INVENTION

Prostate cancer is the most prevalent cancer in men. Diagnosis is basedon a tissue biopsy, microscopic examination of a small section of theprostate. When the biopsy contains cancerous cells the therapeuticoptions generally include radio- and chemotherapies, hormone treatmentsas well as surgical removal of the prostate gland. It is well known thatthese treatments are frequently devastating for the patient andnotwithstanding these therapies mortality from prostate cancer is high.

There is an urgent need for agents that prevent the advancement ofprostate cancer. Indeed, of the approximately one million prostatebiopsies performed in this country every year, by far the majority arenegative. Yet these men suffer a range of symptoms associated withelevated PSA levels and benign prostatic hyperplasia affecting urinaryand sexual function. These large groups of men with BPH (BenignProstatic Hyperplasia) or elevated PSA, or with PIN (prostaticintraepithelial neoplasia), but with negative biopsies, areprecancerous: they are at high risk of developing an aggressive prostatecancer and there have been no effective preventive treatments. Agentsthat forestall or significantly delay the onset of cancer in thispopulation have remarkable therapeutic value.

Another large population of men is diagnosed with prostate cancer, buttheir biopsies reveal a low-grade form of the disease. This group is inthe well-known ‘watchful waiting’ or ‘active surveillance’ category andwhen the rate of increase in their PSA levels is relatively rapid theyare subject to the therapeutic interventions cited above. These largegroups of men—those in watchful waiting and those with shorter PSAdoubling times need agents that subdue their cancers, slow the rate ofPSA increase, and prevent the progression of their cancers to a highergrade and more aggressive form. Such an agent would fill a major vacuumin healthcare for men.

A third population of men undergo one of the traditional therapies inwhich case their recovery is assessed by monitoring the rate of changeor the doubling time of their PSA levels which serve as an indicator ofhow rapidly their cancer is likely to return. An agent that is effectivein reducing PSA levels and in moderating PSA doubling times could beused in combination with traditional therapies to help prevent therecurrence of prostate cancer thus significantly extending the life ofthis population of cancer patients.

SUMMARY OF THE INVENTION

The present invention includes compositions and methods for alleviatingor avoiding the onset of the symptoms associated with prostate cancer,for treating low-grade prostate cancers and preventing their progressionto more advanced cancers, and for decreasing PSA doubling times and thuspreventing or delaying metastatic prostate cancer. The compositionincludes a mixture of flavopereirine and alstonine. Flavopereirine andalstonine are present in defined ratios relative to each other. Theflavopereirine and alstonine may be present as plant extracts. Theextract containing flavopereirine is derived from Pao Pereira and theextract containing alstonine is derived from Rauwolfia Vomitoria. Theextract of flavopereirine to the extract of alstonine ratio is about3-4:1 and is based on the active ingredients being in extract form, i.e.Pao Pereira extract to Rauwolfia Vomitoria extract. Alternatively theflavopereirine and alstonine may be present in purified chemical form,i.e. a combination of synthetic flavopereirine and synthetic alstonine.It is expected that the ratio will vary somewhat depending on the purityof the active ingredients in the case of the active ingredients being inextract form. The amounts are sufficient to induce apoptosis anddisruption of the cell cycle and DNA damage response pathways inprostate cancer cells.

Any conventional modes of administration for compounds of this type areenvisioned. Currently, oral administration is contemplated. Typicalmodes of this type would include tablets and capsules. These could bepackaged in kit form with written instructions to facilitate a treatmentregimen.

The population envisioned for treatment with the flavopereirine andalstonine mixture falls into three groups: 1) those subjects thought tobe susceptible to prostate cancer; 2) those subjects with low-gradeforms of prostate cancer with both low and high PSA doubling times; and3) those subjects combining treatment with traditional medical therapies(specifically surgery, radiation, and cryo- and/or chemotherapy)Population number one could include such high-risk groups as maleshaving a high PSA count, symptoms of BPH (Benign Prostatic Hyperplasia)or with PIN (prostatic intraepithelial neoplasia). This group will havenegative biopsies and would be considered precancerous. The amount ofthe mixture is sufficient to alleviate the symptoms of BPH (BenignProstatic Hyperplasia) or to lower elevated PSA levels over time.Population number two will have been diagnosed with prostate cancer andthus will be likely to have positive biopsies, but have non-aggressivetumors characterized by lower Gleason scores. This population willinclude men with relatively low rates of PSA increase as well as menwith higher rates of PSA increase. The amount of the mixture issufficient to slow the progression of prostate cancer to a moreaggressive stage and to lower PSA levels and PSA doubling times.Population number three will have or will be undergoing some form ofprostate cancer therapy (surgery, radiation, cryo- or chemotherapy).Used in combination with these therapies the amount of the mixture issufficient to lower PSA doubling times.

The invention also includes a novel combination of these two extracts inrelative amounts to prevent prostate cancer or alleviate symptoms ofBPH, especially in precancerous men including men diagnosed withprostatic intraepithelial neoplasia (PIN). The extracts exert beneficialeffects without the side effects normally associated with anti-cancertreatments. e Precancerous cell that are becoming cancerous areselectively killed by the mixture and the normal cells are leftunaffected.

The invention also includes a method of preventing prostatecarcinogenesis in patients having a precancerous precursor of prostateadenocarcinoma, elevated PSA, and Benign Prostatic Hyperplasia, but whodo not as yet have prostate cancer. The invention further includes amethod for reducing PSA levels and/or alleviating the symptoms of BPH.

The invention also includes a method of treating men with localized(non-metastatic) prostate cancer to prevent the development ofhigher-grade tumors and more aggressive disease. The cancer cells areselectively killed by the mixture and the normal cells are leftunaffected.

The invention also includes a method of treating men with both long andshort PSA doubling times and of lengthening the PSA doubling times. Thistreatment will help maintain the ‘watchful waiting’ status of men withlong PSA doubling times and help prevent the onset of more aggressive ormetastatic disease in men with short PSA doubling times.

The invention further provides a method of administering to anindividual an orally effective dosage of the extracts in capsule ortablet form.

The invention provides a method for suppression or inhibition ofprostate carcinogenesis, reduction of PSA levels, and reduction of thesymptoms of BPH by administration of the preventive agent that consistsof a combination of natural extracts derived from Pao Pereira andRauwolfia Vomitoria.

The invention provides a mechanism of action for the preventive agent byinduction of apoptosis and disruption of cell cycle and DNA damageresponse pathways in cancer cells. At optimal concentrations thismechanism is selective for cancer cells and it also contributes to theeffectiveness of the preventive agent in reducing PSA levels and thesymptoms of BPH.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the results of LNCaP cell growth analyses followingtreatment with Rauwolfia and Pao extracts.

FIG. 2 shows the induction of apoptosis in LNCaP cells by both Rauwolfiaand Pao extracts after 24 hours in terms of PARP cleavage and Caspase-3activity.

FIG. 3 shows cell cycle analysis by FACS.

FIG. 4 (A and B) shows the results of a prostate cell tumor growthexperiment.

FIG. 5 shows the results for the TUNEL staining assay.

FIG. 6 shows the corresponding statistical data for the TUNEL stainingassay.

FIG. 7 shows the experimental results illustrating the suppression ofLNCap tumor xenograft growth by Rauwolfia.

DESCRIPTION OF THE INVENTION Overview

The present invention builds on the scientific research of M. Beljanskiwho first developed these extracts and demonstrated their selectiveactivity against cancer cells in laboratory experiments and in animalstudies. Beljanski provided evidence that the toxicity of thesecompounds is based on their interaction with the destabilized DNA thathe found to be a primary characteristic of cancer cells.

The work described herein supports the correlation of the extent of DNAdamage in prostate cells with cancer of the prostate; this also supportsthe specific diagnosis of a precancerous condition of the prostate. Alsodescribed is work demonstrating the mechanism of action involvesinduction of apoptosis and disruption of the cell cycle and DNA damageresponse pathways in prostate cancer cells. This is shown both in tissueculture and in an animal xenograft model. Uncovering the differentialmechanisms of action of flavopereirine and alstonine supports theconcept of using a specific combination of the extracts to obtain anenhanced therapeutic effect.

The classification of an intermediate or precancerous stage of prostateDNA damage indicated that these compounds would exert their toxic effectspecifically on precancerous cells that are in transition to a cancerousstate thereby preventing cancer in high-risk men and answering theurgent need of a very large patient population. These compounds alsoexert their toxic effect on cancer cells thereby preventing advancementto a higher grade and more aggressive form of prostate cancer.

Discovery of the Anti-Cancer Activity of Pao Pereira and RauwolfiaVomitoria Extracts.

The extract from Pao Pereira is prepared according to the protocol inU.S. Pat. No. 5,519,028. The extract from Rauwolfia Vomitoria isprepared according to the procedure in European Pat. No. EP-A-0 059 817.This procedure includes a partial purification step that eliminatestoxic compounds (e.g. reserpine) from the product.

The biochemical activity of these two extracts was first analyzed in anin vitro assay called the Oncotest (U.S. Pat. No. 4,264,729). This testwas originally designed to screen compounds for their carcinogenicpotential and is based on differences in the secondary structure of DNAsisolated from cancer cells and from normal cells (Beljanski, IRCS Med.Sci. 1979. 7:476). Relative to purified normal DNA, purified cancer cellDNA exhibits greater chromicity (absorption of UV light) and greateractivity when used as a template for in vitro DNA synthesis (Beljanskiet al., Expl. Cell. Biol., 1981.49:220-231. This indicates that thehydrogen bonds in the double helix of cancer DNA are reproduciblydisrupted resulting in openings or loops that increase UV absorption andenhance enzymatic activity in vitro. This fundamental difference instructure, the greater strand separation characteristic of cancer DNA,was referred to as DNA destabilization (Beljanski, Proceedings of theinternational seminar: Traditional Medicine: a Challenge to the 21^(st)Century. 1992. Oxford and IBH Publishing Co.)

When known carcinogens are added to the Oncotest they promote a smallincrease in DNA synthesis with normal templates, but a much higherincrease (5 to 10 fold) in DNA synthesis with cancer templates(Beljanski et al. Third NCI-EORTC Symposium on new drugs in cancertherapy. 1981. Institut Bordet, Bruxelles). Such compounds are thoughtto act by further opening the already comparatively relaxed structure ofthe cancer DNA thereby enhancing the access of enzymes to the templateand enabling more DNA to be synthesized.

When the Pao and Rauwolfia extracts are used in the Oncotest they havethe opposite affect of a carcinogen: chromicity is diminished and DNAsynthesis from cancer DNA templates is inhibited (Beljanski, Expl. Cell.Biol. 1982.50: 79-87). Biochemical analysis of the extracts, includingDNA binding assays, identified the active compounds as flavopereirine(in Pao Pereira) and alstonine (in Rauwolfia Vomitoria). Since theinhibitory effects shown by the extracts and their respective compoundsis specific for cancer DNA, since there is no effect on normal DNA, andsince very few of the hundreds of compounds that were screened gavethese results, these extracts and the corresponding active compoundswere further assessed for their potential as anti-cancer agents.

The extracts were subjected to a long series of tests to examine theireffect on cultured cancer cells (Beljanski et al., IRCS Med. Sci. 1984.12:587-588), on animals with various kinds of cancer (Beljanski et al.,Oncology, 1986. 43:198-203), and ultimately in numerous human casestudies. The extracts showed several consistent and noteworthyproperties. First, at optimal concentrations, they stopped theproliferation of cancer cell lines maintained in the laboratory, whilesparing healthy cells. They were toxic to cancer cells in mice, but didno harm to the mice. They have proven to have anti-cancer effects on arange of human malignancies, but have shown no significant side effects.The activity of these extracts is selective to cancer DNA, to cancercells, and to the tumors of organisms with cancer. There is evidencethat this selectivity is due, in part, to their preferential entry intocancer cells because of changes that occur in the outer membranes ofthese cells (Beljanski et al. International Journal of Oncology, 1996.8:1143-1148). In the course of these experiments, the activity of thePao Pereira extract (flavopereirine) (Beljanski, Genetics and Mol.Biol., 2000. 23, 1:29-33) and the Rauwolfia Vomitoria extract(alstonine) (unpublished data) were shown to inhibit the multiplicationof a human prostate cancer cell line (PC3) to a similar extent. Thisresult is significant in the context of the present invention because itdemonstrates that both of the active compounds in the mixture areeffective against cancer cells like PC-3 that do not respond tohormones.

For the Oncotest, cancer DNA was isolated from a range of canceroustissues or cells including breast, lung, ovary, neurocarcinoma, Ehrlichascites tumor cells, KB and HeLa and L cells. Normal DNA was isolatedfrom healthy breast, lung, ovary, brain, primary kidney, Vero cells, orSIRC cells in culture. Cell lines exposed to Pao Pereira extractincluded: u251, CCF-STTG-1 SW 1088 and C6 (brain); LoVo, CaCo-2 (colon);Sk-Hep 1 (liver); A498 (kidney); G-361 (skin); Es 2, Sw 626 (ovary);ZR-75-1, MCF-7 (breast); MIA PaCa2 (pancreas); PC3 (prostate); and TT(thymus). Normal cell lines tested with Pao Pereira include CRL 1656(brain); CCD-18Co (colon); Clone 9 (liver); NRK-49F (kidney); andCCD-97Sk (skin). A summary of anecdotal information concerning the useof the extracts for treating human cancer patients can be found inBeljanski, Mirko Beljanski ou La Chronique d'une “Fatwa” Scientifique,2001, EVI Liberty Corp.

DNA Destabilization in Cancer Cells

Recent studies by Malins et al. (Cancer related changes in prostate DNAas men age and early identification of metastasis in primary prostatetumors. Proc. Natl. Acad. Sci. 2003 April 29 100(9): 5401-5406) usingFourier Transform Mass Spectroscopy have shown that the DNA in prostatecancer cells is indeed different from the DNA of normal prostate cells.Malins et al. refer to the difference as an increase in disorder in thecancer DNA relative to normal DNA and these changes correlate with ageand apparently with exposure to carcinogens. Malins et al were also ableto reproducibly detect an interim stage of disorder reflecting aprecancerous condition of the prostate (Malins et al. Models of DNAstructure achieve almost perfect discrimination between normal prostate,benign prostatic hyperplasia (BPH), and adenocarcinoma and have a highpotential for predicting BPH and prostate cancer. Proc. Natl. Acad. Sci.1997 January 7; 94 (1): 259-264). This precancerous condition appears tocorrespond to the clinical condition known as prostatic intraepithelialneoplasia (PIN).

Malins et al. report that the presence of carcinogens in the prostateleads to the production of free radicals that physically damage the DNAyielding ‘potentially billions of altered structures.’ Many of thesealterations will affect the hydrogen bonds that stabilize the DNAduplex. This could produce the openings in the helix structure reportedby Beljanski.

Novel Data Supporting the Application of Pao Pereira and RauwolfiaExtracts for the Prevention of Prostate Cancer

1. Induction of Apoptosis: The Primary Mechanism of Action ofFlavopereirine

Flavopereirine and alstonine are structurally related alkaloids of thebeta-carboline class. There is evidence that they enter the relativelyopen strands of the DNA in cancer cells and intercalate between adjacentnucleotides where their presence apparently interferes with themetabolism of DNA (Calvez, Flavopereirine is an intercalating agent fornon-supercoiled DNA, Cancer Detection and Prevention 1998; 22(Supplement1)). In effect, the destabilized DNA of the cancer cell is a target ofthese alkaloids and the end result is cell death. The toxicity forcancer cells was confirmed in new experiments showing that both the Paoand Rauwolfia extracts inhibit the growth of LNCaP prostate cancer cellsin tissue culture (FIG. 1). Moreover, these extracts exhibitedsignificantly higher toxicity for LNCaP cells than for a primary cultureof normal prostate cells (Data not shown).

Our recent experiments have also shown that the primary mechanism ofaction of flavopereirine is the induction of apoptosis whereas alstoninehas a relatively minor effect in the induction of this pathway Both thePao Pereira extract and the Rauwolfia Vomitoria extract triggeredapoptosis in the prostate cancer cell line LNCaP as determined bycleavage of PARP and induction of Caspase-3 [FIG. 2], but the effect ofthe Rauwolfia extract was considerably less than the effect of to Paoextract.

Apoptosis—also called programmed cell death—is a built-in biochemicalpathway that triggers a cell to commit suicide. This pathway is normallyused during development (e.g. in the development of the nervous system),by the immune system to protect the body from altered cells (e.g. cellsinfected by a virus), and as a cellular reaction to DNA damage (e.g. ifnot destroyed these cells can become cancerous). In many cancers,including many prostate cancers, the mechanism for inducing apoptosis isdefective so cells that should normally be committing suicide surviveand grow to form a tumor. Numerous cancer therapies actually function byinducing apoptosis in certain types of cancer cells.

The induction of apoptosis is a central theme in the prostate cancerresearch community and the fact that the Pao Pereira and (to asignificantly lower degree Rauwolfia Vomitoria) extracts exhibit thiseffect makes them extremely attractive as potential anti-prostate canceragents.

2. Potential for Synergistic Action of Flavopereirine and AlstonineBased on Differential Effects on the Apoptotic Pathway, on the CellCycle, and on the DNA Damage Response Pathway

Although both extracts induce apoptosis and are toxic for prostatecancer cells, analysis of their action in the cell cycle by fluorescenceactivated cell sorting revealed different effects. As shown in FIG. 3,the Pao Pereira extract causes accumulation of LNCaP cells in SubG0whereas Rauwolfia Vomitoria hinder progression of the cells from G1 toS. This result highlights a difference in the mechanism of action of thetwo extracts whose biochemical basis remains to be determined, though itis presumably based on the difference in the physical structures offlavopereirine and alstonine. One possibility is that they may induceapoptosis by different pathways (e.g. p53-dependent and p53-independent)and another is that they may function with somewhat different kinetics.Regardless of the reason, this difference in activity has therapeuticvalue: combinations of the two extracts have the potential to act incomplementary fashion, that is, they are potentially synergistic intheir anti-cancer effects. Combination therapies are often used incancer treatment and are widely considered to offer significantadvantages over the use of a single anti-cancer agent alone.

To further explore the effect of these two extracts on prostate cancercells a series of microarray assays were performed using arrayscontaining three different sets of human genes belonging to threedifferent biochemical pathways: apoptosis, cell cycle, and DNA damage.In each case that RNAs for the assays were isolated from LNCaP cellsthat had been exposed to the extracts for 24 hours. The data obtainedfrom these microarray experiments are referred to collectively as TableI. See Annex. Note that real time PCR was used to spot check thevalidity of the changes observed in these assays

While the process of analyzing these data and synthesizing coherentdescriptions of the effect of the Pao and Rauwolfia extracts on thesecomplex pathways is ongoing and will require additional experiments,several observations can be made highlighting the fact that the twoextracts induce different responses in all three microarrays. Forexample, in the DNA damage response array the Pao extract has apotentially significant effect on the expression of fourteen (14) genes(shown in bold) whereas the Rauwolfia extract has a potentiallysignificant response on thirty (30) transcripts. Closer inspectionreveals that both extracts have significant effects on the expression ofjust four common genes (ATM, DDIT3, ERCC1, and PDCD8). Even moreremarkable is that of the four genes affected by both extracts only twoare affected in the same way: DDIT3 and ERCC1 are both up-regulated.Analysis of the two other microarrays also reveals divergent results forthe Pao Pereira and Rauwolfia Vomitoria extracts strongly substantiatingthe use of these extracts in combination in order to obtain theircomplementary effects.

3. Flavopereirine and Alstonine are Active Against Prostate CancerTumors in a Xenograft Animal Model

The ability of human tumors to grow as xenografts in athymic mice (e.g.,Balb/c, nu/nu) provides a useful in vivo model for studying thebiological response to therapies for human cancers. Since the firstsuccessful xenotransplantation of human tumors into athymic mice, manydifferent human tumor cell lines (e.g., mammary, lung, genitourinary,gastro-intestinal, head and neck, glioblastoma, bone, and malignantmelanomas) have been transplanted and successfully grown in nude mice.The use of this system enabled analysis of the activity of the compoundsof the present invention on a prostate cancer xenograft of LNCaP cellsin a mammal—a better model for humans than cell-based assays. A numberof tests may be used to determine the level of activity, specificity andeffect of the compounds and in this case tumor volume andimmunohistochemistry were the endpoints.

A graph of the results of the xenograft experiment using the Pao Pereiraextract, with measurement of tumor size (cubic millimeters) plottedagainst time (days), are shown if FIG. 4A. Daily doses of 10 and 20mg/kg of Pao Pereira extract caused a statistically significantreduction in tumor volume compared to control animals and to animalstreated with 50 mg/kg per day. The log transformation shown in FIG. 4Bclearly shows that the results fall into two classes. A graph of theresults of the xenograft experiment using the Rauwolfia Vomitoriaextract is shown in FIG. 7. In this case all three doses of the extractcaused statistically significant reduction in tumor volume relative tothe controls [p<0.0001)]. Finally, data from two immunohistochemicalassays using sections of the tumors removed from the animals confirmedthat the mechanism of action of Pao is the induction of apoptosis whichis dosage dependent. The results for the TUNEL staining assay are shownin FIG. 5 and the corresponding statistical data are shown in FIG. 6.Consistent results were obtained in the BrdU staining assay [Data notshown]. Similar immunohistochemistry for the Rauwolfia Vomitoriaexperiment is currently underway.

The dramatic effect of these extracts in reducing tumor volume must alsobe considered in light of the remarkable fact that the health of theanimals in these experiments is not otherwise compromised in any adverseway. For example, the weights of treated and control mice show nosignificant differences throughout the course of the experiment. Theanti-prostate cancer effects of both extracts together with the absenceof negative side effects provide a strong scientific basis for theclaims described in the present invention.

Formulation and Administration

The flavopereirine and alstonine combinations can be formulated aspharmaceutical compositions. The compositions can be included inpharmaceutical kits. Depending on the subject to be treated, the mode ofadministration, and the type of treatment desired—e.g, prevention,prophylaxis, therapy; the flavopereirine and alstonine combinations areformulated in ways consistent with these parameters. A summary of suchtechniques is found in Remington's Pharmaceutical Sciences, latestedition, Mack Publishing Co., Easton, Pa.

In general, for use in treatment or prophylaxis, the mixture offlavopereirine and alstonine may be used alone or in combination withother chemotherapeutic agents compounds. The mixture of flavopereirineand alstonine can be administrated singly or in combination with otherpharmaceutically active components, and in single or multipleadministrations. The mixture formulation may be prepared in a mannersuitable for systemic administration. Systemic formulations includethose designed for injection, e.g. intramuscular, intravenous orsubcutaneous injection, or may be prepared for transdermal,transmucosal, or oral administration. The formulation will generallyinclude a diluent as well as, in some cases, adjuvants, buffers,preservatives and the like. The flavopereirine and alstonine mixture canbe administered also in liposomal compositions or as microemulsionsusing conventional techniques.

If orally administered, the flavopereirine and alstonine combination ofthe invention can be protected from degradation in the stomach using asuitable enteric coating.

The manner of administration and formulation of the mixture useful inthe invention will depend on the nature of the condition, the severityof the condition, the particular subject to be treated, and the judgmentof the practitioner, formulation will depend on mode of administrationAs the mixture of the invention involve small molecules, it isconveniently administered by oral administration by compounding themwith suitable pharmaceutical excipients so as to provide tablets,capsules, syrups, and the like. Suitable formulations for oraladministration may also include minor components such as buffers,flavoring agents and the like. Typically, the amount of activeingredient in the formulations will be in the range of 1%-99% of thetotal formulation, but wide variation is permitted depending on thecarrier. Suitable carriers include sucrose, pectin, magnesium stearate,lactose, peanut oil, olive oil, water, and the like.

The mixture may also be administered by injection, includingintravenous, intramuscular, subcutaneous or intraperitoneal injection.Typical formulations for such use are liquid formulations in isotonicvehicles such as Hank's solution or Ringer's solution.

Suitable alternative formulations also include liposomal formulations,slow-release formulations, and the like.

Any suitable formulation may be used. A compendium of art-knownformulations is found in Remington's Pharmaceutical Sciences, latestedition, Mack Publishing Company, Easton, Pa. Reference to this manualis routine in the art.

The dosages of the compounds of the invention will depend on a number offactors which will vary from patient to patient.

Oral dosage forms include capsules and tablets. Capsule or tablets canbe easily formulated and can be made easy to swallow. Tablets maycontain suitable carriers, binders, lubricants, diluents, disintegratingagents, coloring agents, flavoring agents, flow-inducing agents, ormelting agents. A tablet may be made by compression or molding,optionally with one or more additional ingredients. Compressed tablesmay be prepared by compressing the active ingredient in a free flowingform (e.g., powder, granules) optionally mixed with a binder (e.g.,gelatin, hydroxypropylmethylcellulose), lubricant, inert diluent,preservative, disintegrant (e.g., sodium starch glycolate, cross-linkedcarboxymethyl cellulose) surface-active or dispersing agent. Suitablebinders include starch, gelatin, natural sugars such as glucose orbeta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth, or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes, or the like. Lubricants used in these dosageforms include sodium oleate, sodium stearate, magnesium stearate, sodiumbenzoate, sodium acetate, sodium chloride, or the like. Disintegratorsinclude, for example, starch, methyl cellulose, agar, bentonite, xanthangum, or the like. Molded tablets may be made by molding in a suitablemachine a mixture of the powdered active ingredient moistened with aninert liquid diluent.

The tablets may optionally be coated or scored and may be formulated soas to provide slow- or controlled-release of the active ingredient.Tablets may also optionally be provided with an enteric coating toprovide release in parts of the gut other than the stomach.

As noted above the flavopereirine and alstonine mixture can be includedin a kit form. The mixture would be formulated for use as apharmaceutical. The kits can comprise one or more containers containingthe pharmaceutical form of the mixture, which comprises atherapeutically effective amount of the mixture in unit dosage form.Such kits can further include, if desired, one or more of variousconventional pharmaceutical kit components, such as, for example,containers with one or more pharmaceutically acceptable carriers,additional containers, etc., as will be readily apparent to thoseskilled in the art. Instructions, such as printed instructions forexample, either as inserts or as labels, the instruction indicatingquantities of the components to be administered, guidelines foradministration, and/or guidelines for further mixing with additionalcomponents, can also be included in the kit.

Tables

Project Pao Pereira Apoptosis 100 ug/ml Pao 24 hr/LNCaP Catalog # OligoGEArray ® Human Apoptosis Microarray Density Average Clover Clover OffTotal Arrays 2 Total Groups 2 Background Empty spots: positions 117,114, 116 Normalization Selected genes: positions 126 Array Name Capoptosis light.tif T apoptosis.tif Assigned Group Group 1 Group 2Position UniGene RefSeq # Symbol Group 2 Group 1 Group 2/Group 1 1Hs.546292 NM_002954 RPS27A 1.462 1.029 1.422 2 Hs.431048 NM_005157 ABL10.130 0.201 0.648 3 Hs.525622 NM_005163 AKT1 0.494 0.607 0.814 4Hs.552567 NM_001160 APAF1 0.019 −0.003 −6.013 5 Hs.370254 NM_004322 BAD0.024 0.027 0.896 6 Hs.377484 NM_004323 BAG1 0.040 0.084 0.472 7Hs.523309 NM_004281 BAG3 0.137 0.117 1.172 8 Hs.194726 NM_004874 BAG40.006 −0.011 −0.491 9 Hs.485139 NM_001188 BAK1 0.187 0.269 0.695 10Hs.159428 NM_004324 BAX 0.026 −0.002 −16.620 11 Hs.549082 NM_003921BCL10 0.016 −0.001 −27.393 12 Hs.150749 NM_000633 BCL2 0.016 −0.002−8.129 13 Hs.227817 NM_004049 BCL2A1 0.012 −0.007 −1.853 14 Hs.516966NM_138578 BCL2L1 0.094 0.139 0.680 15 Hs.283672 NM_020396 BCL2L10 0.013−0.009 −1.464 16 Hs.469658 NM_006538 BCL2L11 0.010 −0.009 −1.046 17Hs.289052 NM_052842 BCL2L12 0.124 0.325 0.383 18 Hs.546359 NM_015367BCL2L13 0.131 0.204 0.642 19 Hs.410026 NM_004050 BCL2L2 0.015 0.00125.098 20 Hs.486542 NM_014739 BCLAF1 0.021 0.029 0.726 21 Hs.435556NM_016561 BFAR 0.004 −0.003 −1.284 22 Hs.474150 NM_001196 BID 0.1280.129 0.993 23 Hs.475055 NM_001197 BIK 0.031 0.006 5.098 24 Hs.519374NM_004536 BIRC1 0.002 −0.013 −0.174 25 Hs.503704 NM_001166 BIRC2 0.0460.066 0.698 26 Hs.127799 NM_001165 BIRC3 0.013 −0.012 −1.136 27Hs.356076 NM_001167 BIRC4 0.006 0.001 6.777 28 Hs.514527 NM_001168 BIRC50.610 0.941 0.648 29 Hs.150107 NM_016252 BIRC6 0.008 −0.003 −2.277 30Hs.256126 NM_022161 BIRC7 0.005 −0.011 −0.441 31 Hs.348263 NM_033341BIRC8 0.011 −0.012 −0.912 32 Hs.145726 NM_001205 BNIP1 0.039 0.007 5.72833 Hs.283454 NM_004330 BNIP2 0.016 0.010 1.541 34 Hs.79428 NM_004052BNIP3 1.091 0.942 1.159 35 Hs.131226 NM_004331 BNIP3L 0.139 0.163 0.85336 Hs.293753 NM_032515 BOK 0.071 0.067 1.064 37 Hs.490366 NM_004333 BRAF0.014 0.001 13.023 38 Hs.57973 NM_014550 CARD10 0.022 0.003 7.058 39Hs.300355 NM_032415 CARD11 0.005 −0.000 −11.164 40 Hs.23248 NM_021209CARD12 0.004 −0.015 −0.266 41 Hs.550529 NM_024110 CARD14 0.008 −0.007−1.258 42 Hs.135201 NM_022162 CARD15 0.011 0.007 1.576 43 Hs.405153NM_006092 CARD4 0.007 −0.008 −0.791 44 Hs.200242 NM_032587 CARD6 0.0050.003 1.868 45 Hs.446146 NM_014959 CARD8 0.002 −0.001 −2.515 46Hs.271815 NM_022352 CARD9 0.013 −0.004 −3.092 47 Hs.2490 NM_033292 CASP10.012 −0.004 −2.743 48 Hs.5353 NM_001230 CASP10 0.016 −0.006 −2.506 49Hs.248226 NM_012114 CASP14 0.011 −0.009 −1.273 50 Hs.368982 NM_001224CASP2 0.123 0.212 0.581 51 Hs.141125 NM_004346 CASP3 0.016 −0.003 −4.95952 Hs.138378 NM_001225 CASP4 0.137 0.189 0.725 53 Hs.213327 NM_004347CASP5 0.008 −0.003 −2.563 54 Hs.3280 NM_001226 CASP6 0.020 0.012 1.69655 Hs.9216 NM_001227 CASP7 0.030 0.027 1.115 56 Hs.369736 NM_001228CASP8 −0.000 −0.008 0.027 57 Hs.329502 NM_001229 CASP9 0.049 0.068 0.72558 Hs.390736 NM_003879 CFLAR 0.009 −0.005 −1.687 59 Hs.249129 NM_001279CIDEA 0.013 0.004 3.523 60 Hs.448590 NM_014430 CIDEB 0.013 0.009 1.44661 Hs.38533 NM_003805 CRADD 0.115 0.176 0.652 62 Hs.380277 NM_004938DAPK1 0.006 0.001 6.290 63 Hs.237886 NM_014326 DAPK2 0.021 0.004 5.06164 Hs.484782 NM_004401 DFFA 0.040 0.025 1.578 65 Hs.133089 NM_004402DFFB 0.040 0.082 0.490 66 Hs.86131 NM_003824 FADD 0.167 0.241 0.690 67Hs.80409 NM_001924 GADD45A 0.177 0.129 1.377 68 Hs.248114 NM_000514 GDNF0.003 0.007 0.431 69 Hs.87247 NM_003806 HRK 0.005 −0.004 −1.276 70Hs.20573 NM_000875 IGF1R −0.002 0.003 −0.701 71 Hs.36 NM_000595 LTA0.005 0.004 1.105 72 Hs.1116 NM_002342 LTBR 0.786 0.751 1.046 73Hs.532826 NM_021960 MCL1 0.459 0.605 0.758 74 Hs.513667 NM_003946 NOL30.246 0.317 0.776 75 Hs.499094 NM_013258 PYCARD 0.011 0.005 2.128 76Hs.519842 NM_003804 RIPK1 0.005 0.002 2.240 77 Hs.103755 NM_003821 RIPK20.072 0.149 0.485 78 Hs.241570 NM_000594 TNF 0.004 0.004 0.807 79Hs.401745 NM_003844 TNFRSF10A 0.031 0.023 1.376 80 Hs.521456 NM_003842TNFRSF10B 0.985 0.921 1.070 81 Hs.119684 NM_003841 TNFRSF10C 0.016 0.0081.991 82 Hs.213467 NM_003840 TNFRSF10D 0.006 0.007 0.927 83 Hs.81791NM_002546 TNFRSF11B 0.024 −0.000 −67.929 84 Hs.355899 NM_016639TNFRSF12A 0.020 0.029 0.692 85 Hs.512898 NM_003820 TNFRSF14 0.009 −0.002−4.653 86 Hs.149168 NM_018647 TNFRSF19 0.003 −0.005 −0.515 87 Hs.279594NM_001065 TNFRSF1A 0.107 0.180 0.595 88 Hs.256278 NM_001066 TNFRSF1B0.006 −0.001 −4.329 89 Hs.443577 NM_014452 TNFRSF21 0.012 −0.007 −1.67690 Hs.462529 NM_003790 TNFRSF25 0.032 0.034 0.950 91 Hs.472860 NM_001250CD40 0.002 −0.007 −0.228 92 Hs.244139 NM_152877 FAS 0.006 −0.009 −0.67393 Hs.434878 NM_003823 TNFRSF6B 0.050 0.082 0.613 94 Hs.355307 NM_001242TNFRSF7 0.006 −0.003 −2.164 95 Hs.193418 NM_001561 TNFRSF9 0.005 0.0017.460 96 Hs.478275 NM_003810 TNFSF10 0.005 −0.002 −2.889 97 Hs.129708NM_003807 TNFSF14 −0.001 0.001 −1.310 98 Hs.248197 NM_005092 TNFSF180.006 −0.002 −2.723 99 Hs.652 NM_000074 CD40LG 0.003 −0.005 −0.555 100Hs.2007 NM_000639 FASLG 0.002 −0.005 −0.368 101 Hs.501497 NM_001252TNFSF7 0.003 0.004 0.754 102 Hs.494901 NM_001244 TNFSF8 0.006 −0.007−0.823 103 Hs.1524 NM_003811 TNFSF9 0.003 −0.004 −0.658 104 Hs.408312NM_000546 TP53 0.009 0.026 0.368 105 Hs.523968 NM_005426 TP53BP2 0.0070.006 1.239 106 Hs.192132 NM_005427 TP73 0.000 −0.006 −0.073 107Hs.137569 NM_003722 TP73L 0.004 −0.005 −0.808 108 Hs.460996 NM_003789TRADD 0.205 0.246 0.835 109 Hs.531251 NM_005658 TRAF1 0.002 −0.000−5.206 110 Hs.522506 NM_021138 TRAF2 0.001 0.016 0.078 111 Hs.510528NM_003300 TRAF3 0.026 0.029 0.894 112 Hs.8375 NM_004295 TRAF4 0.4340.614 0.707 113 Hs.523930 NM_004619 TRAF5 −0.002 −0.005 0.356 114 Blank0.001 −0.002 −0.346 115 N/A L08752 PUC18 0.000 −0.002 −0.209 116 Blank0.001 0.001 1.087 117 Blank −0.002 0.001 −3.216 118 N/A N/A AS1R2 0.000−0.007 −0.016 119 N/A N/A AS1R1 0.004 0.002 1.532 120 N/A N/A AS1 0.013−0.005 −2.927 121 Hs.544577 NM_002046 GAPD 0.878 1.014 0.866 122Hs.534255 NM_004048 B2M 0.642 0.645 0.994 123 Hs.509736 NM_007355 HSPCB0.975 1.006 0.969 124 Hs.509736 NM_007355 HSPCB 0.934 1.039 0.899 125Hs.520640 NM_001101 ACTB 1.008 1.030 0.979 126 Hs.520640 NM_001101 ACTB1.000 1.000 1.000 127 N/A N/A BAS2C 0.055 0.106 0.517 128 N/A N/A BAS2C0.751 0.922 0.815 * As default, Group 1 is used as control group.

Project Rauwolfia Apoptosis 500 ug/ml 24 hr LNCaP Catalog # OligoGEArray ® Human Apoptosis Microarray Density Average Clover Clover OffTotal Arrays 2 Total Groups 2 Background Empty spots: positions 117,114, 116 Normalization Selected genes: positions 1, 121, 123, 124, 125,126 Array Name Rauwolfia Apoptosis Rauwolfia Apoptosis Rx.tif AssignedGroup Group 1 Group 2 Position UniGene RefSeq # Symbol Group 2 Group 1Group 2/Group 1 1 Hs.546292 NM_002954 RPS27A 0.4721 0.4813 0.9809 2Hs.431048 NM_005157 ABL1 −0.1908 −0.0298 6.4113 3 Hs.525622 NM_005163AKT1 −0.0315 0.1583 −0.1989 4 Hs.552567 NM_001160 APAF1 0.3442 −0.0376−9.1443 5 Hs.370254 NM_004322 BAD 0.0596 0.0600 0.9930 6 Hs.377484NM_004323 BAG1 −0.0272 0.1709 −0.1590 7 Hs.523309 NM_004281 BAG3 0.04210.1204 0.3493 8 Hs.194726 NM_004874 BAG4 −0.1620 0.1170 −1.3841 9Hs.485139 NM_001188 BAK1 −0.0838 0.0916 −0.9154 10 Hs.159428 NM_004324BAX −0.2322 0.0343 −6.7676 11 Hs.549082 NM_003921 BCL10 −0.1444 0.0383−3.7758 12 Hs.150749 NM_000633 BCL2 −0.0527 0.0706 −0.7466 13 Hs.227817NM_004049 BCL2A1 −0.0477 0.0874 −0.5461 14 Hs.516966 NM_138578 BCL2L1−0.0831 0.1577 −0.5271 15 Hs.283672 NM_020396 BCL2L10 −0.0871 0.1131−0.7704 16 Hs.469658 NM_006538 BCL2L11 −0.1348 0.1165 −1.1572 17Hs.289052 NM_052842 BCL2L12 −0.0735 −0.0072 10.2457 18 Hs.546359NM_015367 BCL2L13 −0.1448 0.0797 −1.8154 19 Hs.410026 NM_004050 BCL2L2−0.1769 −0.0185 9.5777 20 Hs.486542 NM_014739 BCLAF1 0.0460 0.02721.6914 21 Hs.435556 NM_016561 BFAR −0.0530 0.0711 −0.7457 22 Hs.474150NM_001196 BID −0.0215 0.1373 −0.1569 23 Hs.475055 NM_001197 BIK −0.15440.1039 −1.4857 24 Hs.519374 NM_004536 BIRC1 −0.1686 0.0748 −2.2556 25Hs.503704 NM_001166 BIRC2 −0.0490 0.0285 −1.7181 26 Hs.127799 NM_001165BIRC3 −0.1521 0.0251 −6.0525 27 Hs.356076 NM_001167 BIRC4 −0.1325 0.0703−1.8852 28 Hs.514527 NM_001168 BIRC5 −0.0861 0.1853 −0.4648 29 Hs.150107NM_016252 BIRC6 −0.1136 0.0687 −1.6536 30 Hs.256126 NM_022161 BIRC7−0.1444 0.1063 −1.3592 31 Hs.348263 NM_033341 BIRC8 −0.1464 0.0876−1.6711 32 Hs.145726 NM_001205 BNIP1 −0.1315 0.0661 −1.9900 33 Hs.283454NM_004330 BNIP2 −0.0136 0.0356 −0.3812 34 Hs.79428 NM_004052 BNIP30.1415 0.2796 0.5059 35 Hs.131226 NM_004331 BNIP3L 0.0013 0.0934 0.014236 Hs.293753 NM_032515 BOK 0.0394 0.0417 0.9461 37 Hs.490366 NM_004333BRAF −0.0590 0.0616 −0.9569 38 Hs.57973 NM_014550 CARD10 −0.0769 0.0960−0.8004 39 Hs.300355 NM_032415 CARD11 −0.0676 0.0818 −0.8257 40 Hs.23248NM_021209 CARD12 −0.0497 0.0446 −1.1153 41 Hs.550529 NM_024110 CARD140.0066 −0.0080 −0.8317 42 Hs.135201 NM_022162 CARD15 0.0133 0.02460.5387 43 Hs.405153 NM_006092 CARD4 0.0616 0.0028 21.9971 44 Hs.200242NM_032587 CARD6 0.2282 −0.0227 −10.0675 45 Hs.446146 NM_014959 CARD80.1491 −0.0048 −30.9637 46 Hs.271815 NM_022352 CARD9 0.0212 0.01831.1589 47 Hs.2490 NM_033292 CASP1 0.0374 0.1018 0.3677 48 Hs.5353NM_001230 CASP10 0.0046 0.1286 0.0361 49 Hs.248226 NM_012114 CASP140.0361 0.0033 10.8554 50 Hs.368982 NM_001224 CASP2 0.1027 0.0795 1.292051 Hs.141125 NM_004346 CASP3 0.1252 −0.0479 −2.6152 52 Hs.138378NM_001225 CASP4 0.0924 −0.0382 −2.4217 53 Hs.213327 NM_004347 CASP50.0398 −0.0941 −0.4225 54 Hs.3280 NM_001226 CASP6 −0.0096 −0.0607 0.158155 Hs.9216 NM_001227 CASP7 0.0248 0.0128 1.9441 56 Hs.369736 NM_001228CASP8 0.0490 0.0608 0.8059 57 Hs.329502 NM_001229 CASP9 0.2067 0.013015.8490 58 Hs.390736 NM_003879 CFLAR 0.1444 0.0028 51.5632 59 Hs.249129NM_001279 CIDEA 0.0633 −0.0014 −45.1769 60 Hs.448590 NM_014430 CIDEB0.1517 −0.0411 −3.6957 61 Hs.38533 NM_003805 CRADD 0.0583 −0.0594−0.9809 62 Hs.380277 NM_004938 DAPK1 0.0189 −0.0321 −0.5878 63 Hs.237886NM_014326 DAPK2 0.0388 0.0217 1.7854 64 Hs.484782 NM_004401 DFFA 0.09610.0776 1.2373 65 Hs.133089 NM_004402 DFFB 0.1020 −0.0014 −72.8507 66Hs.86131 NM_003824 FADD 0.2524 −0.0384 −6.5689 67 Hs.80409 NM_001924GADD45A 0.1941 −0.0453 −4.2895 68 Hs.248114 NM_000514 GDNF 0.1362−0.0652 −2.0878 69 Hs.87247 NM_003806 HRK −0.0560 −0.0825 0.6782 70Hs.20573 NM_000875 IGF1R 0.0086 −0.0573 −0.1502 71 Hs.36 NM_000595 LTA−0.0205 −0.0166 1.2349 72 Hs.1116 NM_002342 LTBR 0.0699 0.0950 0.7360 73Hs.532826 NM_021960 MCL1 0.1424 −0.0064 −22.2920 74 Hs.513667 NM_003946NOL3 0.1030 −0.0206 −5.0084 75 Hs.499094 NM_013258 PYCARD 0.0729 −0.0544−1.3386 76 Hs.519842 NM_003804 RIPK1 0.0099 −0.0713 −0.1395 77 Hs.103755NM_003821 RIPK2 0.1348 0.0201 6.6968 78 Hs.241570 NM_000594 TNF −0.0682−0.0844 0.8087 79 Hs.401745 NM_003844 TNFRSF10A −0.0142 −0.0403 0.353880 Hs.521456 NM_003842 TNFRSF10B 0.2047 0.0582 3.5170 81 Hs.119684NM_003841 TNFRSF10C 0.1375 0.0130 10.5406 82 Hs.213467 NM_003840TNFRSF10D −0.0239 −0.0592 0.4031 83 Hs.81791 NM_002546 TNFRSF11B 0.03310.0049 6.7580 84 Hs.355899 NM_016639 TNFRSF12A −0.0023 −0.0119 0.1948 85Hs.512898 NM_003820 TNFRSF14 −0.0212 −0.0345 0.6147 86 Hs.149168NM_018647 TNFRSF19 −0.0232 −0.0870 0.2665 87 Hs.279594 NM_001065TNFRSF1A 0.0199 0.0256 0.7750 88 Hs.256278 NM_001066 TNFRSF1B −0.04440.0002 −253.5583 89 Hs.443577 NM_014452 TNFRSF21 0.0851 −0.0418 −2.034790 Hs.462529 NM_003790 TNFRSF25 0.0437 −0.0689 −0.6347 91 Hs.472860NM_001250 CD40 0.0179 −0.0883 −0.2025 92 Hs.244139 NM_152877 FAS −0.0106−0.0886 0.1197 93 Hs.434878 NM_003823 TNFRSF6B −0.0480 −0.0846 0.5675 94Hs.355307 NM_001242 TNFRSF7 −0.0411 −0.1004 0.4091 95 Hs.193418NM_001561 TNFRSF9 −0.1070 −0.0552 1.9372 96 Hs.478275 NM_003810 TNFSF10−0.0755 −0.0221 3.4105 97 Hs.129708 NM_003807 TNFSF14 0.0358 −0.0613−0.5839 98 Hs.248197 NM_005092 TNFSF18 0.0354 −0.0707 −0.5012 99 Hs.652NM_000074 CD40LG 0.0457 −0.0321 −1.4230 100 Hs.2007 NM_000639 FASLG0.1474 0.1443 1.0213 101 Hs.501497 NM_001252 TNFSF7 −0.0212 −0.06050.3505 102 Hs.494901 NM_001244 TNFSF8 −0.0338 0.0900 −0.3755 103 Hs.1524NM_003811 TNFSF9 −0.0517 0.0380 −1.3603 104 Hs.408312 NM_000546 TP530.0083 0.1110 0.0746 105 Hs.523968 NM_005426 TP53BP2 0.1159 0.02255.1539 106 Hs.192132 NM_005427 TP73 0.0520 −0.0295 −1.7631 107 Hs.137569NM_003722 TP73L 0.0855 0.0485 1.7624 108 Hs.460996 NM_003789 TRADD0.0480 0.1010 0.4755 109 Hs.531251 NM_005658 TRAF1 −0.0133 −0.07100.1867 110 Hs.522506 NM_021138 TRAF2 −0.0288 −0.0355 0.8110 111Hs.510528 NM_003300 TRAF3 −0.1259 −0.0363 3.4653 112 Hs.8375 NM_004295TRAF4 0.0149 0.0771 0.1933 113 Hs.523930 NM_004619 TRAF5 0.0172 −0.0298−0.5788 114 Blank 0.0500 −0.0547 −0.9143 115 N/A L08752 PUC18 −0.0126−0.0463 0.2719 116 Blank −0.0636 −0.0064 9.9536 117 Blank 0.0136 0.06110.2223 118 N/A N/A AS1R2 −0.0129 0.1307 −0.0989 119 N/A N/A AS1R1 0.26530.1974 1.3443 120 N/A N/A AS1 0.9584 0.8912 1.0754 121 Hs.544577NM_002046 GAPD 0.7006 1.0997 0.6371 122 Hs.534255 NM_004048 B2M 0.28390.1086 2.6134 123 Hs.509736 NM_007355 HSPCB 0.9918 0.8395 1.1815 124Hs.509736 NM_007355 HSPCB 0.8686 0.8668 1.0021 125 Hs.520640 NM_001101ACTB 1.5778 1.2042 1.3103 126 Hs.520640 NM_001101 ACTB 1.3890 1.50860.9208 127 N/A N/A BAS2C 2.1066 1.7055 1.2351 128 N/A N/A BAS2C 2.09961.6617 1.2635 * As default, Group 1 is used as control group.

Project Pao Cell Cycle 100 ug/ml 24 hr LNCaP Catalog # Oligo GEArray ®Human Cell Cycle Microarray Density Average Clover Clover Off TotalArrays 2 Total Groups 2 Background Empty spots: positions 117, 114, 116Normalization Selected genes: positions 125 Array Name T cell cyclelight.tif T cell cycle light.tif Assigned Group Group 1 Group 2 PositionUniGene RefSeq # Symbol Group 2 Group 1 Group 2/Group 1 1 Hs.546292NM_002954 RPS27A 0.992 0.956 1.039 2 Hs.431048 NM_005157 ABL1 0.0480.078 0.622 3 Hs.533262 NM_013366 ANAPC2 0.010 0.009 1.056 4 Hs.152173NM_013367 ANAPC4 0.049 0.106 0.467 5 Hs.7101 NM_016237 ANAPC5 0.1180.281 0.422 6 Hs.194695 NM_004675 ARHI 0.002 −0.000 −6.380 7 Hs.435561NM_000051 ATM 0.029 0.042 0.682 8 Hs.271791 NM_001184 ATR 0.015 0.0310.493 9 Hs.159428 NM_004324 BAX 0.007 0.004 1.659 10 Hs.370292 NM_016567BCCIP 0.012 0.020 0.605 11 Hs.150749 NM_000633 BCL2 0.004 0.001 6.347 12Hs.514527 NM_001168 BIRC5 0.224 0.806 0.278 13 Hs.194143 NM_007294 BRCA10.006 0.030 0.182 14 Hs.34012 NM_000059 BRCA2 0.003 0.000 35.636 15Hs.417050 NM_003914 CCNA1 0.002 0.002 1.177 16 Hs.85137 NM_001237 CCNA20.007 0.014 0.476 17 Hs.23960 NM_031966 CCNB1 0.048 0.128 0.371 18Hs.194698 NM_004701 CCNB2 0.008 0.004 1.945 19 Hs.430646 NM_005190 CCNC0.013 0.004 3.493 20 Hs.523852 NM_053056 CCND1 0.007 0.000 16.573 21Hs.376071 NM_001759 CCND2 0.000 −0.000 −2.759 22 Hs.534307 NM_001760CCND3 0.337 0.805 0.419 23 Hs.244723 NM_001238 CCNE1 0.042 0.080 0.52524 Hs.408658 NM_004702 CCNE2 0.007 0.009 0.774 25 Hs.1973 NM_001761 CCNF0.002 0.001 3.677 26 Hs.79101 NM_004060 CCNG1 0.014 0.013 1.033 27Hs.13291 NM_004354 CCNG2 0.057 0.098 0.579 28 Hs.146607 NM_001239 CCNH0.244 0.691 0.353 29 Hs.279906 NM_001240 CCNT1 0.004 −0.000 −14.118 30Hs.292754 NM_001241 CCNT2 0.015 0.013 1.161 31 Hs.374127 NM_003903 CDC160.083 0.140 0.594 32 Hs.334562 NM_001786 CDC2 0.006 0.004 1.365 33Hs.524947 NM_001255 CDC20 0.062 0.362 0.171 34 Hs.1634 NM_001789 CDC25A0.001 −0.003 −0.307 35 Hs.656 NM_001790 CDC25C 0.008 0.021 0.366 36Hs.514997 NM_004359 CDC34 0.031 0.060 0.516 37 Hs.160958 NM_007065 CDC370.020 0.015 1.333 38 Hs.474217 NM_003504 CDC45L 0.003 0.000 37.346 39Hs.405958 NM_001254 CDC6 0.007 0.009 0.793 40 Hs.533573 NM_003503 CDC70.001 0.002 0.569 41 Hs.19192 NM_001798 CDK2 0.045 0.137 0.330 42Hs.95577 NM_000075 CDK4 0.770 0.957 0.805 43 Hs.500015 NM_003885 CDK5R10.004 −0.002 −2.473 44 Hs.158460 NM_003936 CDK5R2 0.002 −0.000 −4.346 45Hs.435952 NM_016408 CDK5RAP1 0.006 0.010 0.576 46 Hs.20157 NM_176096CDK5RAP3 0.001 0.000 7.638 47 Hs.119882 NM_001259 CDK6 0.003 0.001 5.75048 Hs.184298 NM_001799 CDK7 0.037 0.036 1.036 49 Hs.382306 NM_001260CDK8 0.022 0.003 8.117 50 Hs.370771 NM_000389 CDKN1A 0.912 0.865 1.05451 Hs.238990 NM_004064 CDKN1B 0.041 0.043 0.956 52 Hs.106070 NM_000076CDKN1C 0.001 −0.002 −0.481 53 Hs.421349 NM_058195 CDKN2A 0.012 0.0260.452 54 Hs.72901 NM_004936 CDKN2B 0.006 0.005 1.094 55 Hs.525324NM_078626 CDKN2C 0.021 0.068 0.312 56 Hs.435051 NM_001800 CDKN2D 0.0190.045 0.424 57 Hs.84113 NM_005192 CDKN3 0.025 0.018 1.406 58 Hs.24529NM_001274 CHEK1 0.007 0.002 3.101 59 Hs.291363 NM_007194 CHEK2 0.0020.003 0.593 60 Hs.374378 NM_001826 CKS1B 0.045 0.123 0.369 61 Hs.83758NM_001827 CKS2 0.330 0.679 0.486 62 Hs.146806 NM_003592 CUL1 0.399 0.5380.743 63 Hs.82919 NM_003591 CUL2 0.010 0.015 0.662 64 Hs.372286NM_003590 CUL3 0.008 0.000 20.679 65 Hs.339735 NM_003589 CUL4A 0.0430.022 1.923 66 Hs.440320 NM_003478 CUL5 0.016 0.002 9.800 67 Hs.443960NM_004399 DDX11 0.008 0.015 0.525 68 Hs.211463 NM_004945 DNM2 0.0030.002 1.762 69 Hs.96055 NM_005225 E2F1 0.100 0.278 0.359 70 Hs.194333NM_004091 E2F2 0.015 0.045 0.322 71 Hs.269408 NM_001949 E2F3 0.004 0.0015.082 72 Hs.108371 NM_001950 E2F4 0.008 0.004 2.180 73 Hs.445758NM_001951 E2F5 0.008 0.000 98.497 74 Hs.135465 NM_001952 E2F6 0.001−0.001 −0.743 75 Hs.80409 NM_001924 GADD45A 0.075 0.032 2.310 76Hs.523510 NM_005316 GTF2H1 0.024 0.013 1.858 77 Hs.386189 NM_016426GTSE1 0.006 0.007 0.828 78 Hs.26663 NM_016323 HERC5 0.001 −0.001 −1.46679 Hs.152983 NM_004507 HUS1 0.004 −0.000 −28.200 80 Hs.300559 NM_014708KNTC1 0.005 0.004 1.207 81 Hs.252712 NM_002266 KPNA2 0.180 0.346 0.52082 Hs.533185 NM_002358 MAD2L1 0.026 0.035 0.737 83 Hs.19400 NM_006341MAD2L2 0.118 0.154 0.767 84 Hs.477481 NM_004526 MCM2 0.377 0.563 0.66985 Hs.179565 NM_002388 MCM3 0.007 0.007 1.025 86 Hs.460184 NM_005914MCM4 0.003 0.002 2.108 87 Hs.517582 NM_006739 MCM5 0.009 0.016 0.600 88Hs.444118 NM_005915 MCM6 0.010 0.008 1.202 89 Hs.438720 NM_005916 MCM70.142 0.345 0.411 90 Hs.80976 NM_002417 MKI67 0.006 0.032 0.194 91Hs.509523 NM_002431 MNAT1 0.003 0.002 1.406 92 Hs.20555 NM_005590 MRE11A0.001 0.004 0.350 93 Hs.492208 NM_002485 NBS1 0.003 0.002 1.834 94Hs.147433 NM_182649 PCNA 0.514 0.744 0.691 95 Hs.77783 NM_182687 PKMYT10.029 0.144 0.201 96 Hs.531879 NM_002853 RAD1 0.017 0.011 1.554 97Hs.16184 NM_002873 RAD17 0.006 0.001 9.384 98 Hs.242635 NM_005732 RAD500.005 0.001 4.031 99 Hs.446554 NM_002875 RAD51 0.003 −0.001 −2.643 100Hs.240457 NM_004584 RAD9A 0.010 0.020 0.489 101 Hs.408528 NM_000321 RB10.032 0.010 3.188 102 Hs.546282 NM_002894 RBBP8 0.010 0.011 0.890 103Hs.207745 NM_002895 RBL1 0.008 0.005 1.414 104 Hs.513609 NM_005611 RBL20.023 0.018 1.307 105 Hs.507866 NM_014059 RGC32 0.005 −0.003 −1.854 106Hs.487540 NM_002947 RPA3 0.002 0.000 28.121 107 Hs.269898 NM_013376SERTAD1 0.009 0.006 1.665 108 Hs.23348 NM_005983 SKP2 −0.000 −0.0010.039 109 Hs.81424 NM_003352 SUMO1 0.027 0.027 1.012 110 Hs.79353NM_007111 TFDP1 0.035 0.038 0.901 111 Hs.379018 NM_006286 TFDP2 0.0080.006 1.346 112 Hs.408312 NM_000546 TP53 0.016 0.018 0.914 113 Hs.406693NM_003334 UBE1 0.061 0.044 1.381 114 Blank −0.000 0.001 −0.240 115 N/AL08752 PUC18 −0.001 −0.001 0.911 116 Blank −0.000 −0.001 0.190 117 Blank0.000 0.001 0.555 118 N/A N/A AS1R2 0.006 0.002 3.958 119 N/A N/A AS1R1−0.000 0.004 −0.110 120 N/A N/A AS1 0.003 0.004 0.627 121 Hs.544577NM_002046 GAPD 0.911 0.990 0.920 122 Hs.534255 NM_004048 B2M 0.333 0.4870.684 123 Hs.509736 NM_007355 HSPCB 0.981 0.917 1.070 124 Hs.509736NM_007355 HSPCB 0.987 0.964 1.024 125 Hs.520640 NM_001101 ACTB 1.0001.000 1.000 126 Hs.520640 NM_001101 ACTB 0.980 0.967 1.013 127 N/A N/ABAS2C 0.032 0.093 0.347 128 N/A N/A BAS2C 0.669 0.841 0.795 * Asdefault, Group 1 is used as control group.

Project Rauwolfia Cell Cycle 500 ug/ml 24 hr LNCaP Catalog # OligoGEArray ® Human Cell Cycle Microarray Density Average Clover Clover OffTotal Arrays 2 Total Groups 2 Background Empty spots: positions 117,114, 116 Normalization Selected genes: positions 1, 121, 122, 123, 124,125, 126 Array Name Rauwolfia Rx Cell Cycle.tif Assigned Group Group 1Group 2 Position UniGene RefSeq # Symbol Group 2 Group 1 Group 2/Group 11 Hs.546292 NM_002954 RPS27A 0.9708 0.7410 1.3102 2 Hs.431048 NM_005157ABL1 0.0617 0.0930 0.6641 3 Hs.533262 NM_013366 ANAPC2 0.3128 0.34050.9186 4 Hs.152173 NM_013367 ANAPC4 0.0785 0.0452 1.7375 5 Hs.7101NM_016237 ANAPC5 0.0074 −0.0339 −0.2169 6 Hs.194695 NM_004675 ARHI0.0092 −0.0952 −0.0969 7 Hs.435561 NM_000051 ATM 0.5787 0.5695 1.0162 8Hs.271791 NM_001184 ATR 0.1767 −0.0030 −57.9743 9 Hs.159428 NM_004324BAX 0.0493 0.0381 1.2941 10 Hs.370292 NM_016567 BCCIP 0.1229 0.03243.7965 11 Hs.150749 NM_000633 BCL2 0.1167 0.0552 2.1130 12 Hs.514527NM_001168 BIRC5 0.0996 0.1983 0.5024 13 Hs.194143 NM_007294 BRCA1 0.12450.1423 0.8747 14 Hs.34012 NM_000059 BRCA2 0.0328 −0.0721 −0.4555 15Hs.417050 NM_003914 CCNA1 0.2733 −0.0522 −5.2368 16 Hs.85137 NM_001237CCNA2 0.2059 −0.0202 −10.1973 17 Hs.23960 NM_031966 CCNB1 0.0664 0.05201.2757 18 Hs.194698 NM_004701 CCNB2 0.1124 0.0495 2.2690 19 Hs.430646NM_005190 CCNC 0.0888 0.0834 1.0648 20 Hs.523852 NM_053056 CCND1 0.01360.6031 0.0225 21 Hs.376071 NM_001759 CCND2 0.0042 −0.0152 −0.2801 22Hs.534307 NM_001760 CCND3 0.1037 0.1890 0.5487 23 Hs.244723 NM_001238CCNE1 0.1121 −0.0053 −21.0110 24 Hs.408658 NM_004702 CCNE2 0.0241−0.0277 −0.8701 25 Hs.1973 NM_001761 CCNF 0.0869 0.0509 1.7073 26Hs.79101 NM_004060 CCNG1 0.1559 0.1108 1.4070 27 Hs.13291 NM_004354CCNG2 0.0962 0.1149 0.8374 28 Hs.146607 NM_001239 CCNH 0.0272 0.10420.2615 29 Hs.279906 NM_001240 CCNT1 −0.0501 −0.0081 6.2066 30 Hs.292754NM_001241 CCNT2 0.0285 −0.0394 −0.7230 31 Hs.374127 NM_003903 CDC160.1177 −0.0072 −16.4273 32 Hs.334562 NM_001786 CDC2 −0.0191 −0.04280.4450 33 Hs.524947 NM_001255 CDC20 0.1136 0.0948 1.1987 34 Hs.1634NM_001789 CDC25A 0.0962 0.1126 0.8544 35 Hs.656 NM_001790 CDC25C 0.06860.0795 0.8628 36 Hs.514997 NM_004359 CDC34 0.0282 −0.0332 −0.8480 37Hs.160958 NM_007065 CDC37 −0.0100 −0.0202 0.4976 38 Hs.474217 NM_003504CDC45L −0.0501 −0.1244 0.4029 39 Hs.405958 NM_001254 CDC6 0.0204 −0.1619−0.1260 40 Hs.533573 NM_003503 CDC7 −0.0085 0.0013 −6.5566 41 Hs.19192NM_001798 CDK2 0.2109 0.2500 0.8435 42 Hs.95577 NM_000075 CDK4 0.44820.7110 0.6304 43 Hs.500015 NM_003885 CDK5R1 0.1462 0.1259 1.1618 44Hs.158460 NM_003936 CDK5R2 0.0869 0.0280 3.0991 45 Hs.435952 NM_016408CDK5RAP1 0.0738 −0.0846 −0.8724 46 Hs.20157 NM_176096 CDK5RAP3 0.07420.0242 3.0702 47 Hs.119882 NM_001259 CDK6 −0.0271 0.0811 −0.3347 48Hs.184298 NM_001799 CDK7 0.0055 0.0248 0.2210 49 Hs.382306 NM_001260CDK8 0.1003 0.1291 0.7767 50 Hs.370771 NM_000389 CDKN1A 2.1099 0.49464.2662 51 Hs.238990 NM_004064 CDKN1B 0.1525 0.1284 1.1875 52 Hs.106070NM_000076 CDKN1C 0.0493 0.0280 1.7582 53 Hs.421349 NM_058195 CDKN2A0.0307 0.0098 3.1434 54 Hs.72901 NM_004936 CDKN2B 0.0515 0.0575 0.894855 Hs.525324 NM_078626 CDKN2C 0.1229 0.0891 1.3802 56 Hs.435051NM_001800 CDKN2D 0.0639 0.0626 1.0215 57 Hs.84113 NM_005192 CDKN3 0.06670.0790 0.8442 58 Hs.24529 NM_001274 CHEK1 0.0863 0.1007 0.8565 59Hs.291363 NM_007194 CHEK2 0.1260 0.0820 1.5374 60 Hs.374378 NM_001826CKS1B 0.0978 0.0578 1.6929 61 Hs.83758 NM_001827 CKS2 0.1111 0.13390.8301 62 Hs.146806 NM_003592 CUL1 0.1422 0.1199 1.1857 63 Hs.82919NM_003591 CUL2 0.1726 0.0644 2.6816 64 Hs.372286 NM_003590 CUL3 0.14470.0637 2.2715 65 Hs.339735 NM_003589 CUL4A 0.1267 0.1076 1.1774 66Hs.440320 NM_003478 CUL5 0.4051 0.4319 0.9378 67 Hs.443960 NM_004399DDX11 0.1369 0.1250 1.0957 68 Hs.211463 NM_004945 DNM2 0.1456 0.008417.3744 69 Hs.96055 NM_005225 E2F1 0.1357 0.3227 0.4205 70 Hs.194333NM_004091 E2F2 0.0244 0.0223 1.0949 71 Hs.269408 NM_001949 E2F3 0.13630.0443 3.0789 72 Hs.108371 NM_001950 E2F4 0.1388 0.0482 2.8821 73Hs.445758 NM_001951 E2F5 −0.0265 0.0454 −0.5839 74 Hs.135465 NM_001952E2F6 0.0316 0.0079 3.9864 75 Hs.80409 NM_001924 GADD45A 0.1465 0.04753.0873 76 Hs.523510 NM_005316 GTF2H1 0.0515 0.0123 4.1961 77 Hs.386189NM_016426 GTSE1 0.0878 0.0397 2.2125 78 Hs.26663 NM_016323 HERC5 0.02070.0461 0.4494 79 Hs.152983 NM_004507 HUS1 0.0947 0.0226 4.1973 80Hs.300559 NM_014708 KNTC1 0.0748 0.0148 5.0589 81 Hs.252712 NM_002266KPNA2 0.0014 0.1711 0.0085 82 Hs.533185 NM_002358 MAD2L1 0.0608 0.08980.6773 83 Hs.19400 NM_006341 MAD2L2 0.0154 0.1848 0.0835 84 Hs.477481NM_004526 MCM2 0.0791 0.1746 0.4533 85 Hs.179565 NM_002388 MCM3 0.0018−0.0213 −0.0825 86 Hs.460184 NM_005914 MCM4 0.0201 −0.0149 −1.3454 87Hs.517582 NM_006739 MCM5 0.1204 0.0107 11.2919 88 Hs.444118 NM_005915MCM6 0.0735 0.0216 3.3983 89 Hs.438720 NM_005916 MCM7 0.0617 0.09890.6241 90 Hs.80976 NM_002417 MKI67 0.1220 0.0134 9.0981 91 Hs.509523NM_002431 MNAT1 0.0192 0.0200 0.9562 92 Hs.20555 NM_005590 MRE11A 0.0372−0.0350 −1.0608 93 Hs.492208 NM_002485 NBS1 0.0732 −0.0394 −1.8589 94Hs.147433 NM_182649 PCNA 0.1419 0.0591 2.3998 95 Hs.77783 NM_182687PKMYT1 0.1034 0.0472 2.1880 96 Hs.531879 NM_002853 RAD1 0.1034 0.03083.3579 97 Hs.16184 NM_002873 RAD17 0.0331 0.0141 2.3512 98 Hs.242635NM_005732 RAD50 0.0540 −0.0636 −0.8481 99 Hs.446554 NM_002875 RAD510.0058 −0.0325 −0.1783 100 Hs.240457 NM_004584 RAD9A −0.0085 −0.01880.4513 101 Hs.408528 NM_000321 RB1 0.3510 0.2383 1.4727 102 Hs.546282NM_002894 RBBP8 0.1354 0.0173 7.8265 103 Hs.207745 NM_002895 RBL1 0.06390.0749 0.8532 104 Hs.513609 NM_005611 RBL2 0.1556 0.0706 2.2048 105Hs.507866 NM_014059 RGC32 −0.0243 0.0751 −0.3240 106 Hs.487540 NM_002947RPA3 0.0275 −0.0382 −0.7203 107 Hs.269898 NM_013376 SERTAD1 0.0272−0.0819 −0.3326 108 Hs.23348 NM_005983 SKP2 0.0533 −0.1075 −0.4961 109Hs.81424 NM_003352 SUMO1 0.1121 0.0468 2.3954 110 Hs.79353 NM_007111TFDP1 0.0645 −0.0069 −9.3060 111 Hs.379018 NM_006286 TFDP2 0.1382 0.05042.7391 112 Hs.408312 NM_000546 TP53 0.3084 0.1853 1.6645 113 Hs.406693NM_003334 UBE1 0.0257 0.1261 0.2037 114 Blank 0.0428 0.0150 2.8497 115N/A L08752 PUC18 0.1037 0.0852 1.2171 116 Blank −0.0228 −0.0449 0.5077117 Blank −0.0200 0.0299 −0.6692 118 N/A N/A AS1R2 0.0319 0.0395 0.8082119 N/A N/A AS1R1 0.3081 0.1775 1.7356 120 N/A N/A AS1 0.8820 0.70961.2428 121 Hs.544577 NM_002046 GAPD 0.8549 1.3506 0.6330 122 Hs.534255NM_004048 B2M 0.2289 0.1485 1.5413 123 Hs.509736 NM_007355 HSPCB 0.71791.0587 0.6781 124 Hs.509736 NM_007355 HSPCB 0.7975 0.9087 0.8776 125Hs.520640 NM_001101 ACTB 1.6684 1.4658 1.1382 126 Hs.520640 NM_001101ACTB 1.7616 1.3268 1.3277 127 N/A N/A BAS2C 2.1083 1.4589 1.4451 128 N/AN/A BAS2C 2.0903 1.4185 1.4736 * As default, Group 1 is used as controlgroup.

Project Pao Pereira DNA damage 100 ug/ml Pao, 24 hr, LNCaP Catalog #Oligo GEArray ® Human DNA Damage Signaling Pathway Microarray DensityAverage Clover Clover Off Total Arrays 2 Total Groups 2 Background Emptyspots: positions 117, 116 Normalization Selected genes: positions 1,121, 123, 124, 125, 126 Array Name Pao Rx DNA damage scan dark.tifAssigned Group Group 1 Group 2 Position UniGene RefSeq # Symbol Group 2Group 1 Group 2/Group 1 1 Hs.546292 NM_002954 RPS27A 0.776 0.898 0.864 2Hs.431048 NM_005157 ABL1 −0.000 0.057 −0.006 3 Hs.518804 NM_198889ANKRD17 0.004 0.010 0.467 4 Hs.73722 NM_080649 APEX1 0.009 0.113 0.080 5Hs.154149 NM_014481 APEX2 −0.044 −0.017 2.615 6 Hs.435561 NM_000051 ATM0.335 0.181 1.857 7 Hs.271791 NM_001184 ATR −0.015 −0.031 0.501 8Hs.533526 NM_000489 ATRX 0.024 0.009 2.752 9 Hs.194143 NM_007294 BRCA11.480 0.030 49.394 10 Hs.34012 NM_000059 BRCA2 0.020 0.034 0.600 11Hs.519162 NM_006763 BTG2 0.088 −0.029 −3.060 12 Hs.146607 NM_001239 CCNH0.065 0.093 0.695 13 Hs.184298 NM_001799 CDK7 0.019 −0.025 −0.775 14Hs.24529 NM_001274 CHEK1 −0.002 −0.034 0.047 15 Hs.291363 NM_007194CHEK2 −0.005 −0.054 0.100 16 Hs.135471 NM_006384 CIB1 0.317 0.826 0.38417 Hs.249129 NM_001279 CIDEA −0.014 0.033 −0.426 18 Hs.448590 NM_014430CIDEB −0.006 0.054 −0.112 19 Hs.435237 NM_000082 ERCC8 0.065 −0.010−6.217 20 Hs.151573 NM_004075 CRY1 0.061 −0.025 −2.399 21 Hs.532491NM_021117 CRY2 0.030 −0.084 −0.357 22 Hs.290758 NM_001923 DDB1 −0.005−0.063 0.076 23 Hs.446564 NM_000107 DDB2 −0.022 −0.072 0.309 24Hs.505777 NM_004083 DDIT3 0.876 0.035 24.790 25 Hs.339396 NM_007068 DMC1−0.047 0.041 −1.139 26 Hs.435981 NM_001983 ERCCI 0.044 0.132 0.333 27Hs.487294 NM_000400 ERCC2 0.055 −0.036 −1.514 28 Hs.469872 NM_000122ERCC3 0.161 −0.024 −6.716 29 Hs.460019 NM_005236 ERCC4 0.028 −0.075−0.368 30 Hs.258429 NM_000123 ERCC5 −0.069 −0.050 1.378 31 Hs.498248NM_130398 EXO1 −0.066 −0.098 0.675 32 Hs.434873 NM_004629 FANCG −0.044−0.082 0.528 33 Hs.409065 NM_004111 FEN1 0.083 0.074 1.113 34 Hs.292493NM_001469 G22P1 1.067 0.974 1.095 35 Hs.80409 NM_001924 GADD45A 0.3790.047 8.015 36 Hs.9701 NM_006705 GADD45G 0.193 −0.030 −6.488 37 Hs.86161NM_002066 GML 0.068 −0.059 −1.138 38 Hs.523510 NM_005316 GTF2H1 0.024−0.016 −1.519 39 Hs.191356 NM_001515 GTF2H2 0.107 0.161 0.661 40Hs.355348 NM_001516 GTF2H3 −0.108 −0.011 9.991 41 Hs.386189 NM_016426GTSE1 0.044 −0.004 −10.361 42 Hs.152983 NM_004507 HUS1 0.099 −0.003−36.939 43 Hs.503048 NM_002180 IGHMBP2 0.105 −0.024 −4.407 44 Hs.17253NM_054111 IHPK3 0.019 −0.065 −0.295 45 Hs.523875 NM_001567 INPPL1 0.007−0.067 −0.111 46 Hs.61188 NM_033276 KUB3 −0.033 −0.030 1.104 47 Hs.1770NM_000234 LIG1 0.028 −0.033 −0.851 48 Hs.100299 NM_002311 LIG3 −0.039−0.031 1.263 49 Hs.166091 NM_002312 LIG4 0.052 0.006 8.808 50 Hs.463978NM_002758 MAP2K6 0.046 0.111 0.413 51 Hs.432642 NM_002969 MAPK12 0.126−0.026 −4.904 52 Hs.35947 NM_003925 MBD4 0.036 −0.053 −0.694 53Hs.195364 NM_000249 MLH1 0.090 0.030 2.994 54 Hs.279843 NM_014381 MLH3−0.175 −0.034 5.158 55 Hs.509523 NM_002431 MNAT1 −0.044 0.008 −5.234 56Hs.459596 NM_002434 MPG −0.197 0.154 −1.278 57 Hs.20555 NM_005590 MRE11A0.125 0.017 7.281 58 Hs.156519 NM_000251 MSH2 0.126 0.072 1.757 59Hs.280987 NM_002439 MSH3 0.124 0.011 10.811 60 Hs.216639 NM_002440 MSH40.114 −0.050 −2.272 61 Hs.371225 NM_002441 MSH5 0.315 0.167 1.888 62Hs.445052 NM_000179 MSH6 0.082 0.041 1.997 63 Hs.271353 NM_012222 MUTYH0.042 0.035 1.181 64 Hs.396494 NM_018177 N4BP2 −0.032 0.047 −0.675 65Hs.492208 NM_002485 NBS1 0.070 0.008 8.919 66 Hs.66196 NM_002528 NTHL10.098 0.083 1.181 67 Hs.534331 NM_002452 NUDT1 0.173 0.039 4.398 68Hs.380271 NM_002542 OGG1 0.112 −0.003 −41.960 69 Hs.20930 NM_020418PCBP4 0.124 −0.054 −2.315 70 Hs.147433 NM_182649 PCNA 0.253 0.154 1.64671 Hs.424932 NM_004208 PDCD8 0.125 0.020 6.383 72 Hs.111749 NM_000534PMS1 0.099 0.057 1.733 73 Hs.487470 NM_000535 PMS2 0.084 −0.006 −13.30174 Hs.278468 D38500 PMS2L4 0.110 0.033 3.311 75 Hs.278467 NM_005395PMS2L9 0.089 −0.001 −147.706 76 Hs.78016 NM_007254 PNKP 0.056 −0.035−1.608 77 Hs.76556 NM_014330 PPP1R15A 0.025 −0.051 −0.486 78 Hs.491682NM_006904 PRKDC 0.034 0.001 44.059 79 Hs.531879 NM_002853 RAD1 0.057−0.026 −2.234 80 Hs.16184 NM_002873 RAD17 −0.038 0.027 −1.397 81Hs.375684 NM_020165 RAD18 0.155 −0.045 −3.469 82 Hs.81848 NM_006265RAD21 0.094 0.023 4.143 83 Hs.440960 NM_005053 RAD23A 0.095 0.001157.765 84 Hs.521640 NM_002874 RAD23B 0.206 0.048 4.307 85 Hs.242635NM_005732 RAD50 −0.024 −0.110 0.219 86 Hs.446554 NM_002875 RAD51 0.028−0.037 −0.776 87 Hs.412587 NM_058216 RAD51C 0.063 −0.004 −14.315 88Hs.172587 NM_133509 RAD51L1 −0.146 0.018 −8.225 89 Hs.125244 NM_002878RAD51L3 0.069 −0.039 −1.759 90 Hs.552577 NM_002879 RAD52 0.064 −0.025−2.530 91 Hs.523220 NM_003579 RAD54L 0.004 −0.001 −5.353 92 Hs.240457NM_004584 RAD9A 0.088 −0.050 −1.754 93 Hs.546282 NM_002894 RBBP8 −0.031−0.061 0.511 94 Hs.443077 NM_016316 REV1L 0.085 −0.054 −1.572 95Hs.461925 NM_002945 RPA1 0.044 0.041 1.081 96 Hs.487540 NM_002947 RPA3−0.120 −0.012 10.294 97 Hs.408846 NM_022367 SEMA4A 0.030 −0.019 −1.60398 Hs.59554 NM_014454 SESN1 −0.002 −0.028 0.056 99 Hs.211602 NM_006306SMC1L1 −0.047 −0.025 1.911 100 Hs.81424 NM_003352 SUMO1 0.138 0.0158.923 101 Hs.408312 NM_000546 TP53 0.182 0.288 0.631 102 Hs.192132NM_005427 TP73 −0.025 −0.100 0.246 103 Hs.344812 NM_016381 TREX1 −0.012−0.004 3.189 104 Hs.170835 NM_007205 TREX2 −0.089 −0.026 3.446 105Hs.191334 NM_003362 UNG −0.049 −0.057 0.866 106 Hs.3041 NM_021147 UNG2−0.027 −0.031 0.866 107 Hs.288867 NM_000380 XPA −0.003 −0.043 0.059 108Hs.475538 NM_004628 XPC 0.069 −0.037 −1.883 109 Hs.98493 NM_006297 XRCC10.063 −0.034 −1.839 110 Hs.129727 NM_005431 XRCC2 0.020 −0.051 −0.399111 Hs.549075 NM_005432 XRCC3 −0.059 −0.060 0.979 112 Hs.171190NM_003401 XRCC4 0.008 0.082 0.098 113 Hs.388739 NM_021141 XRCC5 0.0860.131 0.659 114 Hs.444451 NM_016653 ZAK 0.062 −0.056 −1.124 115 N/AL08752 PUC18 −0.010 0.002 −6.046 116 Blank 0.030 −0.019 −1.565 117 Blank−0.030 0.019 −1.565 118 N/A N/A AS1R2 0.024 −0.031 −0.772 119 N/A N/AAS1R1 0.034 0.074 0.461 120 N/A N/A AS1 0.372 0.249 1.495 121 Hs.544577NM_002046 GAPD 0.830 0.949 0.874 122 Hs.534255 NM_004048 B2M 0.220 0.0346.413 123 Hs.509736 NM_007355 HSPCB 0.873 0.846 1.031 124 Hs.509736NM_007355 HSPCB 0.856 1.072 0.799 125 Hs.520640 NM_001101 ACTB 1.4001.114 1.257 126 Hs.520640 NM_001101 ACTB 1.265 1.120 1.129 127 N/A N/ABAS2C 2.082 1.110 1.875 128 N/A N/A BAS2C 2.005 1.061 1.889 * Asdefault, Group 1 is used as control group.

Project Rauwolfia DNA damage 500 ug/ml 24 hr LNCaP Catalog # OligoGEArray ® Human DNA Damage Signaling Pathway Microarray Density AverageClover Clover Off Total Arrays 2 Total Groups 2 Background Empty spots:positions 117, 116 Normalization Selected genes: positions 1, 121, 123,124, 125, 126 Array Name Rauw Rx DNA damage scans.tif1 Assigned GroupGroup 1 Group 2 Position UniGene RefSeq # Symbol Group 2 Group 1 Group2/Group 1 1 Hs.546292 NM_002954 RPS27A 0.6354 0.9338 0.6805 2 Hs.431048NM_005157 ABL1 −0.0417 0.0205 −2.0340 3 Hs.518804 NM_198889 ANKRD17−0.0848 −0.0055 15.4607 4 Hs.73722 NM_080649 APEX1 −0.0515 0.1662−0.3097 5 Hs.154149 NM_014481 APEX2 −0.0692 0.0076 −9.1234 6 Hs.435561NM_000051 ATM 0.0617 0.1933 0.3191 7 Hs.271791 NM_001184 ATR −0.0596−0.0426 1.3992 8 Hs.533526 NM_000489 ATRX 0.1284 −0.0299 −4.3005 9Hs.194143 NM_007294 BRCA1 0.1155 0.1689 0.6834 10 Hs.34012 NM_000059BRCA2 −0.0404 0.0103 −3.9150 11 Hs.519162 NM_006763 BTG2 −0.0746 −0.00848.8919 12 Hs.146607 NM_001239 CCNH −0.0194 0.1838 −0.1055 13 Hs.184298NM_001799 CDK7 −0.0500 −0.0158 3.1630 14 Hs.24529 NM_001274 CHEK1−0.0584 −0.0986 0.5919 15 Hs.291363 NM_007194 CHEK2 −0.0361 −0.06240.5774 16 Hs.135471 NM_006384 CIB1 0.6782 0.8983 0.7549 17 Hs.249129NM_001279 CIDEA 0.0004 0.0297 0.0140 18 Hs.448590 NM_014430 CIDEB−0.0294 −0.0242 1.2141 19 Hs.435237 NM_000082 ERCC8 −0.0554 0.0219−2.5261 20 Hs.151573 NM_004075 CRY1 −0.0779 −0.0613 1.2712 21 Hs.532491NM_021117 CRY2 −0.0269 −0.0423 0.6359 22 Hs.290758 NM_001923 DDB1 0.0006−0.0534 −0.0117 23 Hs.446564 NM_000107 DDB2 −0.0008 −0.0607 0.0137 24Hs.505777 NM_004083 DDIT3 0.9845 0.0360 27.3602 25 Hs.339396 NM_007068DMC1 0.0336 −0.0683 −0.4916 26 Hs.435981 NM_001983 ERCC1 0.0759 0.01554.8972 27 Hs.487294 NM_000400 ERCC2 −0.0131 −0.0434 0.3025 28 Hs.469872NM_000122 ERCC3 0.0027 −0.0257 −0.1056 29 Hs.460019 NM_005236 ERCC4−0.0313 −0.0660 0.4737 30 Hs.258429 NM_000123 ERCC5 0.0361 −0.0205−1.7594 31 Hs.498248 NM_130398 EXO1 0.0383 −0.0444 −0.8642 32 Hs.434873NM_004629 FANCG 0.1994 −0.0245 −8.1317 33 Hs.409065 NM_004111 FEN10.0325 0.1731 0.1878 34 Hs.292493 NM_001469 G22P1 0.9926 0.9859 1.006835 Hs.80409 NM_001924 GADD45A 0.0417 0.0545 0.7642 36 Hs.9701 NM_006705GADD45G 0.0694 0.0820 0.8466 37 Hs.86161 NM_002066 GML −0.0117 −0.02110.5521 38 Hs.523510 NM_005316 GTF2H1 0.0296 −0.0150 −1.9721 39 Hs.191356NM_001515 GTF2H2 0.0790 0.1478 0.5344 40 Hs.355348 NM_001516 GTF2H30.2003 −0.0452 −4.4328 41 Hs.386189 NM_016426 GTSE1 0.0411 0.1202 0.341542 Hs.152983 NM_004507 HUS1 0.0671 −0.0718 −0.9346 43 Hs.503048NM_002180 IGHMBP2 0.0085 −0.0340 −0.2510 44 Hs.17253 NM_054111 IHPK30.0227 −0.0366 −0.6202 45 Hs.523875 NM_001567 INPPL1 0.0406 −0.0286−1.4229 46 Hs.61188 NM_033276 KUB3 0.0679 −0.0760 −0.8939 47 Hs.1770NM_000234 LIG1 0.0361 −0.0508 −0.7093 48 Hs.100299 NM_002311 LIG3 0.2926−0.0583 −5.0231 49 Hs.166091 NM_002312 LIG4 0.0996 −0.0568 −1.7539 50Hs.463978 NM_002758 MAP2K6 0.0142 −0.0573 −0.2474 51 Hs.432642 NM_002969MAPK12 −0.0177 −0.0729 0.2429 52 Hs.35947 NM_003925 MBD4 0.0240 −0.0365−0.6572 53 Hs.195364 NM_000249 MLH1 0.0377 0.0497 0.7590 54 Hs.279843NM_014381 MLH3 0.0338 −0.0473 −0.7141 55 Hs.509523 NM_002431 MNAT10.0754 −0.0192 −3.9289 56 Hs.459596 NM_002434 MPG 0.2536 0.0287 8.830557 Hs.20555 NM_005590 MRE11A 0.1284 −0.0755 −1.7000 58 Hs.156519NM_000251 MSH2 0.0652 −0.0415 −1.5730 59 Hs.280987 NM_002439 MSH3 0.0213−0.1175 −0.1810 60 Hs.216639 NM_002440 MSH4 0.0092 −0.0710 −0.1292 61Hs.371225 NM_002441 MSH5 0.1496 0.1609 0.9301 62 Hs.445052 NM_000179MSH6 0.1536 0.0513 2.9933 63 Hs.271353 NM_012222 MUTYH 0.1463 −0.0058−25.1854 64 Hs.396494 NM_018177 N4BP2 0.3633 −0.0276 −13.1648 65Hs.492208 NM_002485 NBS1 0.0648 −0.0266 −2.4344 66 Hs.66196 NM_002528NTHL1 0.0915 −0.0097 −9.4499 67 Hs.534331 NM_002452 NUDT1 0.0163 −0.1196−0.1360 68 Hs.380271 NM_002542 OGG1 0.0125 −0.0127 −0.9809 69 Hs.20930NM_020418 PCBP4 0.0681 −0.0424 −1.6059 70 Hs.147433 NM_182649 PCNA0.1517 0.1196 1.2689 71 Hs.424932 NM_004208 PDCD8 0.1836 −0.0066−27.7528 72 Hs.111749 NM_000534 PMS1 0.3674 −0.0366 −10.0309 73Hs.487470 NM_000535 PMS2 0.1184 −0.0239 −4.9568 74 Hs.278468 D38500PMS2L4 0.0536 −0.0026 −20.7457 75 Hs.278467 NM_005395 PMS2L9 0.0577−0.0126 −4.5867 76 Hs.78016 NM_007254 PNKP 0.0571 −0.0010 −58.9812 77Hs.76556 NM_014330 PPP1R15A 0.0688 0.0079 8.6983 78 Hs.491682 NM_006904PRKDC 0.1077 0.0828 1.3016 79 Hs.531879 NM_002853 RAD1 0.2147 0.012617.0552 80 Hs.16184 NM_002873 RAD17 0.3462 0.0602 5.7514 81 Hs.375684NM_020165 RAD18 0.0844 0.0005 174.3605 82 Hs.81848 NM_006265 RAD210.0554 0.0347 1.5979 83 Hs.440960 NM_005053 RAD23A 0.0796 0.0040 19.735084 Hs.521640 NM_002874 RAD23B 0.1207 0.1649 0.7317 85 Hs.242635NM_005732 RAD50 −0.0102 0.0208 −0.4906 86 Hs.446554 NM_002875 RAD510.1002 0.0224 4.4694 87 Hs.412587 NM_058216 RAD51C 0.3126 0.0616 5.071688 Hs.172587 NM_133509 RAD51L1 0.2734 0.1962 1.3935 89 Hs.125244NM_002878 RAD51L3 0.1080 −0.0608 −1.7746 90 Hs.552577 NM_002879 RAD52−0.0046 −0.0423 0.1085 91 Hs.523220 NM_003579 RAD54L 0.0146 −0.0160−0.9132 92 Hs.240457 NM_004584 RAD9A 0.0523 0.0403 1.2967 93 Hs.546282NM_002894 RBBP8 −0.0088 0.0158 −0.5535 94 Hs.443077 NM_016316 REV1L0.0598 0.0813 0.7355 95 Hs.461925 NM_002945 RPA1 0.3061 0.1349 2.2695 96Hs.487540 NM_002947 RPA3 0.2409 0.0896 2.6902 97 Hs.408846 NM_022367SEMA4A 0.0596 −0.0650 −0.9166 98 Hs.59554 NM_014454 SESN1 0.0046 −0.0878−0.0522 99 Hs.211602 NM_006306 SMC1L1 −0.0294 −0.0452 0.6504 100Hs.81424 NM_003352 SUMO1 0.0892 0.0858 1.0391 101 Hs.408312 NM_000546TP53 0.1676 0.3542 0.4731 102 Hs.192132 NM_005427 TP73 0.0490 0.00568.6719 103 Hs.344812 NM_016381 TREX1 0.1369 0.1105 1.2388 104 Hs.170835NM_007205 TREX2 0.2392 0.0889 2.6909 105 Hs.191334 NM_003362 UNG 0.0117−0.0269 −0.4331 106 Hs.3041 NM_021147 UNG2 −0.0131 −0.0850 0.1544 107Hs.288867 NM_000380 XPA −0.0244 0.0261 −0.9328 108 Hs.475538 NM_004628XPC 0.0031 −0.0494 −0.0633 109 Hs.98493 NM_006297 XRCC1 −0.0210 −0.03130.6724 110 Hs.129727 NM_005431 XRCC2 0.0306 0.0127 2.4033 111 Hs.549075NM_005432 XRCC3 0.2405 0.0118 20.4173 112 Hs.171190 NM_003401 XRCC40.2501 0.0694 3.6044 113 Hs.388739 NM_021141 XRCC5 0.0681 0.0699 0.9754114 Hs.444451 NM_016653 ZAK 0.0117 −0.0035 −3.2876 115 N/A L08752 PUC18−0.0390 0.0203 −1.9168 116 Blank 0.0044 0.0082 0.5318 117 Blank −0.0044−0.0082 0.5318 118 N/A N/A AS1R2 0.0990 0.0157 6.3246 119 N/A N/A AS1R10.2345 0.0536 4.3765 120 N/A N/A AS1 0.3899 0.1944 2.0054 121 Hs.544577NM_002046 GAPD 0.3374 0.9567 0.3527 122 Hs.534255 NM_004048 B2M 0.09770.0687 1.4219 123 Hs.509736 NM_007355 HSPCB 0.9589 1.0138 0.9458 124Hs.509736 NM_007355 HSPCB 0.9947 1.0253 0.9702 125 Hs.520640 NM_001101ACTB 1.5266 1.0338 1.4766 126 Hs.520640 NM_001101 ACTB 1.5470 1.03661.4924 127 N/A N/A BAS2C 1.5366 1.0200 1.5065 128 N/A N/A BAS2C 1.57140.9833 1.5980 * As default, Group 1 is used as control group.

1. A method for treating prostate cancer or delaying its onsetcomprising administering an effective amount of a mixture offlavopereirine and alstonine to a subject suspected of being susceptibleor having precancerous or prostate cancer.
 2. The method of claim 1wherein the flavopereirine and alstonine are separately contained innatural extracts.
 3. The method of claim 2 wherein the natural extractsrespectively are derived from Pao Pereira and Rauwolfia Vomitoria. 4.The method of claim 3 wherein the extract of Pao Pereira containsflavopereirine and the extract of Rauwolfia Vomitoria containsalstonine.
 5. The method of claim 4 are combined in the ratio X:Y. 6.The method of claim 1 wherein the flavopereirine and alstonine aresynthetic.
 7. The method of claim 1 wherein the flavopereirine andalstonine are in a purified form.
 8. The method of claim 1 wherein themixture results from the separate administrations of flavopereirine andalstonine to the subject.
 9. The method of claim 1 wherein the subjectis in a high risk group.
 10. The method of claim 9 wherein the high riskgroup includes subjects with a high PSA count, symptoms of BPH (BenignProstatic Hyperplasia) or symptoms of prostatic intraepithelialneoplasia (PIN).
 11. The method of claim 1 wherein the subject has beendiagnosed with prostate cancer but has non-aggressive tumors asevidenced by lower Gleason scores.
 12. The method of claim 1 wherein thesubject is undergoing cancer therapy including surgery, chemotherapy,cryotherapy or radiation.
 13. The method of claim 1 wherein the amountis sufficient to alleviate the symptoms of BPH (Benign ProstaticHyperplasia) or lower elevated PSA. Levels.
 14. The method of claim 1wherein administration is oral.
 15. The method of claim 14 wherein oraladministration is via a tablet or capsule.
 16. The method of claim 1wherein the effective amount is sufficient to induce apoptosis in cancercells.
 17. A composition for treating prostate cancer comprising aneffective amount of a mixture of flavopereirine and alstonine.
 18. Thecomposition of claim 17 wherein the flavopereirine and alstonine areseparately contained in natural extracts.
 19. The composition of claim17 wherein the effective amount is sufficient to alleviate the symptomsof BPH (Benign Prostatic Hyperplasia) or lower elevated PSA. Levels. 20.The composition of claim 17 is in a form suitable for oraladministration.
 21. The composition of claim 20 wherein the form is atablet or a capsule.
 22. The composition of claim 13 wherein the naturalextracts respectively are derived from Pao Pereira and RauwolfiaVomitoria.
 23. The composition of claim 18 wherein the extract of PaoPereira contains flavopereirine and the extract of Rauwolfia Vomitoriacontains alstonine.
 24. The composition of claim 23 wherein the extractsare combined in the ratio X:Y.
 25. The composition of claim 17 whereinthe effective amount is sufficient to induce apoptosis in cancer cells.26. The composition of claim 17 wherein the flavopereirine and alstonineare synthetic.
 27. The composition of claim 17 wherein theflavopereirine and alstonine are in a purified form.
 28. A kitcomprising the composition of claim 17.